Theoretical and experimental approaches for the initiation and propagation of activity in spatially embedded neuronal cultures

[eng] Spatial embedding and inherited metric constraints are a fundamental trait of biological neuronal circuits. However their role in shaping connectivity and dynamics has been often disregarded, with models of neuronal networks paying much more attention to the distribution of connections in the quest for understanding network's behavior. In this thesis we aim at filling this gap by studying the importance of metric features in the complex connectivity- dynamics-noise interplay that shapes spontaneous neuronal activity. This thesis combines experiments in rat dissociated neuronal cultures with theoretical analyses to better comprehend the relevance of spatial embedding. We developed a new theoretical model grounded on Ising Models to assess metric effects in neuronal cultures' behavior, and in the context of percolation approaches. Once metric effects were settled, we illustrated their relevance in shaping spontaneous activity by perturbing the structural connectivity blueprint of neuronal cultures. This was achieved by patterning the substrate where neurons grow, and by using topographical molds that dictated the connectivity of the network. Next, and since the initiation of bursting activity is governed in great manner by a complex amplification mechanism that involves metric correlations and noise, we focused on the metric-driven amplification of spontaneous single-neuron noise to derive an analytical model that predicts the frequency of bursting events in neuronal cultures. We then further investigated in an experimental context the contribution of noise to the observed activity patterns, and by implementing a moderate electrical stimulation protocol that increases the level of activity noise in cultures. Finally, the latter study was completed with experiments regarding the specific role of inhibition in neuronal networks, to provide a wider understanding of the mechanisms that govern the initiation and propagation of activity fronts in cortical cultures.[cat] L'objectiu d'aquesta tesis és investigar els mecanismes que generen l'activitat espontània i estimulada en xarxes neuronals, més concretament en cultius corticals dissociats, i fent un especial èmfasi en l’efecte de les correlacions mètriques. En aquest marc, l’activitat col·lectiva consisteix en episodis esporàdics de dispars quasi sincronitzats entre totes les neurones del cultiu, anomenats “esclats de xarxa”. Tres elements principals en determinen les característiques: connectivitat entre neurones, dinàmica intrínseca neuronal, i soroll (activacions neuronals aleatòries). La investigació s’ha centrat en cinc línies de recerca: l’estudi de correlacions mètriques en cultius neuronals; el desenvolupament d’un model teòric per descriure i predir l’esclat de xarxa; l’anàlisi de la propagació dels fronts d’activitat experimentals sota pertorbacions estructurals de la connectivitat del cultiu; l’estudi de l’efecte de la inhibició en la iniciació i propagació dels esclats ‘in vitro’; i l’estudi de la resposta experimental dels cultius sota una estimulació elèctrica moderada de baixa freqüència. En la primera línia de recerca hem comprovat que les correlacions mètriques dominen el comportament dinàmic del cultiu, fins al punt d’emmascarar la contribució de la distribució del nombre de connexions. En la segona línia hem desenvolupat un model analític que prediu semi- quantitativament la freqüència dels esclats observada experimentalment. La tercera línia s’ha centrat en l’efecte de pertorbacions estructurals en la connectivitat; la dinàmica resultant ha mostrat una gran riquesa en patrons d’activitat, esclats de xarxa a diferents escales, i propagació altament específica de cada cultiu. La quarta línia de recerca ha demostrat que les xarxes sense inhibició disminueixen la seva freqüència d’esclat respecte a les xarxes control, que la velocitat de propagació de l’activitat incrementa lleugerament quan s’ha bloquejat la inhibició, i que els punts on s’inicien ens esclats varien respecte als controls. I, finalment, la cinquena línia de recerca ha constatat que l’aplicació d’un camp elèctric feble augmenta el soroll d’activitat de la xarxa, generant un increment en la freqüència dels esclats de xarxa

Coupled environmental and demographic fluctuations shape the evolution of cooperative antimicrobial resistance

There is a pressing need to better understand how microbial populations respond to antimicrobial drugs, and to find mechanisms to possibly eradicate antimicrobial-resistant cells. The inactivation of antimicrobials by resistant microbes can often be viewed as a cooperative behavior leading to the coexistence of resistant and sensitive cells in large populations and static environments. This picture is however greatly altered by the fluctuations arising in volatile environments, in which microbial communities commonly evolve. Here, we study the eco-evolutionary dynamics of a population consisting of an antimicrobial resistant strain and microbes sensitive to antimicrobial drugs in a time-fluctuating environment, modeled by a carrying capacity randomly switching between states of abundance and scarcity. We assume that antimicrobial resistance is a shared public good when the number of resistant cells exceeds a certain threshold. Eco-evolutionary dynamics is thus characterized by demographic noise (birth and death events) coupled to environmental fluctuations which can cause population bottlenecks. By combining analytical and computational means, we determine the environmental conditions for the long-lived coexistence and fixation of both strains, and characterize a fluctuation-driven antimicrobial resistance eradication mechanism, where resistant microbes experience bottlenecks leading to extinction. We also discuss the possible applications of our findings to laboratory-controlled experiments.Comment: 19+7 pages, 4+1 figures. Simulation data and codes for all figures are electronically available from the University of Leeds Data Repository. DOI: https://doi.org/10.5518/136

Coexistence of Competing Microbial Strains under Twofold Environmental Variability and Demographic Fluctuations

Microbial populations generally evolve in volatile environments, under conditions fluctuating between harsh and mild, e.g. as the result of sudden changes in toxin concentration or nutrient abundance. Environmental variability thus shapes the population long-time dynamics, notably by influencing the ability of different strains of microorganisms to coexist. Inspired by the evolution of antimicrobial resistance, we study the dynamics of a community consisting of two competing strains subject to twofold environmental variability. The level of toxin varies in time, favouring the growth of one strain under low levels and the other strain when the toxin level is high. We also model time-changing resource abundance by a randomly switching carrying capacity that drives the fluctuating size of the community. While one strain dominates in a static environment, we show that species coexistence is possible in the presence of environmental variability. By computational and analytical means, we determine the environmental conditions under which long-lived coexistence is possible and when it is almost certain. We also determine how the make-up of the coexistence phase and the average abundance of each strain depend on the environmental variability

Characterization of fruit products by capillary zone electrophoresis and liquid chromatography using the compositional profiles of polyphenols. Application to authentication of natural extracts

Capillary zone electrophoresis (CZE) and high performance liquid chromatography (HPLC) were applied to the authentication of fruit products based on the compositional profiles of polyphenols. Various sample treatments were used to maximize the overall recovery of polyphenols or specific fractions, such as phenolic acids or anthocyanins. The resulting CZE and HPLC data were treated with Principal Component Analysis (PCA) showing that samples were mainly clustered according to the fruit of origin, with cranberry- and grape-based products clearly separated in groups. A possible adulterated cranberry extract was analyzed more deeply by high resolution mass spectrometry (HRMS) in order to identify the presence of A-type proanthocyanidins, which are characteristic and more abundant in cranberry-based products. In accordance with PCA interpretation, HRMS results indicated that the suspicious sample was not a cranberry-based product, allowing us to validate and demonstrate the suitability of both CZE- and HPLC-proposed methods for the characterization of fruit-based products

Determination of polyphenolic profiles by liquid chromatography-electrospray-tandem mass spectrometry for the authentication of fruit extracts

Liquid chromatography-electrospray-tandem mass spectrometry (LC-ESI-MS/MS) was applied to the analysis and authentication of fruit-based products and fruit-based pharmaceutical preparations. A Kinetex C18 reversed-phase column under gradient elution with 0.1 % formic acid aqueous solution and methanol mobile phases was used for the simultaneous determination of 26 polyphenols, allowing an acceptable separation in less than 22 min. Instrumental quality parameters such as limits of detection (LOD, values between 12-14 µg/L for 19 of the 26 analyzed polyphenols), linearity (r2 > 0.991), run-to-run and day-to-day precisions (RSD values lower than 9.9 and 13.5 %, respectively), and accuracy (relative errors lower than 8 %) were established. A simple extraction method, consisting of a sample sonication with acetone:water:hydrochloric acid (70:29.9:0.1 v/v/v) and centrifugation, was proposed. Two calibration procedures, external calibration using standards prepared in water and standard addition, were evaluated for polyphenol quantification in several grape and cranberry fruits and processed fruit products. For a 95 % confidence level, no statistical differences were observed between the two calibration methods (p values between 0.06 and 0.95), denoting that external calibration was suitable enough for the quantitative analysis of polyphenols in fruit-based products. The proposed LC-ESI-MS/MS method was then applied to the analysis of polyphenols in 23 grape-based and cranberry-based natural products and pharmaceutical preparations. Polyphenolic concentration data was then analyzed by principal component analysis (PCA) to extract information of the most significant profile data contributing to authentication of natural extracts according to their fruit of origin

Dominance of metric correlations in two-dimensional neuronal cultures described through a Random Field Ising Model

We introduce a novel random field Ising model, grounded on experimental observations, to assess the importance of metric correlations in cortical circuits in vitro. Metric correlations arise from both the finite axonal length and the heterogeneity in the spatial arrangement of neurons. The experiments consider the response of neuronal cultures to an external electric stimulation for a gradually weaker connectivity strength between neurons, and in cultures with different spatial configurations. The model can be analytically solved in the metric-free, mean-field scenario. The presence of metric correlations precipitates a strong deviation from the mean field. Null models of the same networks that preserve the distribution of connections recover the mean field. Our results show that metric-inherited correlations in spatial networks dominate the connectivity blueprint, mask the actual distribution of connections, and may emerge as the asset that shapes network dynamics

Neuropsychiatric profiles and conversion to dementia in mild cognitive impairment, a latent class analysis

Altres ajuts: Fondo Europeo de Desarrollo Regional (FEDER)Altres ajuts: Generalitat de Catalunya. Programa CERCAAltres ajuts: Instituto de Salud Carlos III (CIBERSAM i CIBERNED)Altres ajuts: Fundació "La Caixa"Altres ajuts: Grífols SA (GR@ACE project)Neuropsychiatric symptoms (NPS) have been recently addressed as risk factors of conversion to Alzheimer's disease (AD) and other dementia types in patients diagnosed with Mild Cognitive Impairment (MCI). Our aim was to determine profiles based on the prominent NPS in MCI patients and to explore the predictive value of these profiles on conversion to specific types of dementia. A total of 2137 MCI patients monitored in a memory clinic were included in the study. Four NPS profiles emerged (classes), which were defined by preeminent symptoms: Irritability, Apathy, Anxiety/Depression and Asymptomatic. Irritability and Apathy were predictors of conversion to dementia (HR = 1.43 and 1.56, respectively). Anxiety/depression class showed no risk effect of conversion when compared to Asymptomatic class. Irritability class appeared as the most discriminant neuropsychiatric condition to identify non-AD converters (i.e., frontotemporal dementia, vascular dementia, Parkinson's disease and dementia with Lewy Bodies). The findings revealed that consistent subgroups of MCI patients could be identified among comorbid basal NPS. The preeminent NPS showed to behave differentially on conversion to dementia, beyond AD. Therefore, NPS should be used as early diagnosis facilitators, and should also guide clinicians to detect patients with different illness trajectories in the progression of MCI

Medicamentos y alimentos on line: cuestiones éticas en torno al acceso a alimentos y medicamentos a través de Internet

Este artículo plantea cuestiones éticas relevantes pero aun poco debatidas relativas a la accesibilidad de los alimentos y medicamentos a través de Internet, tomando como punto de partida su incidencia de esta situación en la salud colectiva. El objetivo principal de este trabajo es identificar conflictos bioéticos derivados de la utilización de las tecnologías de la información y la comunicación para adquirir medicamentos y alimentos. El artículo distingue entre los riesgos derivados del acceso fácil a medicamentos y alimentos ilegales en Internet, y las implicaciones bioéticas que plantea la adquisición on line de medicamentos y alimentos a través de portales legales.This article describes the ethical implications that arise when buying food and drugs online. The main goal is to identify relevant bioethical conflicts that derive from using information and communication technologies, but which have been overlooked by academia and legislators. The article distinguishes between the risks that easy access to illegal products entails, and the bioethical implications that arise when buying legal food and drugs online

Food and drugs online: ethics of internet-based access to food and medicines

This article describes the ethical implications that arise when buying food and drugs online. The main goal is to identify relevant bioethical conflicts that derive from using information and communication technologies, but which have been overlooked by academia and legislators. The article distinguishes between the risks that easy access to illegal products entails, and the bioethical implications that arise when buying legal food and drugs online.Este artículo plantea cuestiones éticas relevantes pero aun poco debatidas relativas a la accesibilidad de los alimentos y medicamentos a través de Internet, tomando como punto de partida su incidencia de esta situación en la salud colectiva. El objetivo principal de este trabajo es identificar conflictos bioéticos derivados de la utilización de las tecnologías de la información y la comunicación para adquirir medicamentos y alimentos. El artículo distingue entre los riesgos derivados del acceso fácil a medicamentos y alimentos ilegales en Internet, y las implicaciones bioéticas que plantea la adquisición on line de medicamentos y alimentos a través de portales legales.Peer ReviewedPostprint (published version